Use of hyaluronidase for preventing and treating cardio vascular diseases

ABSTRACT

The invention relates to use of the enzyme hyaluronidase in a dosage of at least 6,000 units per day for prevention and/or treatment of cardiovascular diseases whereby the hyaluronidase is administered intra venam over a period of at least four weeks. Hyaluronidase is particularly suited to treating coronary heart diseases as well as for preventive purposes in patients at risk of heart infarctions.

The present invention relates to the use of hyaluronidase for preventionand treatment of cardiovascular diseases such as coronary heart diseaseas well as other arterioscleroses as well as for prevention ofrestenosis in patients who have already been subjected to a coronaryintervention. Coronary interventions include, for example, stentimplantation, balloon dilatation, rotablation and bypass operations(arterio-occlusive disease treatments).

The present invention furthermore relates to prevention and treatment ofacute cardiac infarction.

Cardiovascular diseases and the disturbance of well-being and theappearance of physical impairments as its sequelae have in recent yearsaffected an increasing number of people. By now numerous medicationshave been developed in order to treat heart diseases. Thesecardiopharmaceuticals include complex compounds like vegetable extractsas well as individual chemical compounds such as those listed in the“2000 Red List”. But none of the currently marketed medications hasshown really satisfactory results in treating cardiovascular diseases.

The application NL8102880 describes the use of 20,000-80,000 IU ofhyaluronidase for treatment of arteriosclerosis, enhanced blood supplyand embolism. The hyaluronidase is in these cases appliedintra-arterially for some 15-30 minutes at intervals of three weeks.

The object of the present invention is to solve the technical problem ofindicating an additional possibility of preventing and treatingcardiovascular diseases in humans or animals.

This problem is solved by the present invention according to patentclaim 1 by using hyaluronidase in a dosage of at least 6,000 units perday for prevention and/or treatment of cardiovascular diseases where thehyaluronidase is administered intra venam over a minimum period of fourweeks, preferably in a bolus such as within ten seconds.

Additional advantageous and preferred embodiments of the invention arethe subject of subclaims.

According to one embodiment of the invention, the hyaluronidase is usedin a dosage of at least 12,000 units per day,

According to a further embodiment of the invention the hyaluronidase isused in a dosage of at least 15,000 units per day. However, it is alsopossible to work with a dosage of more than 100.000 units per day.

According to still another embodiment of the invention the hyaluronidaseis additionally used in combination with camitin and/or CSE inhibitorsand/or nootropics and/or blood supply enhancement agents and/or vitaminsand/or cations.

It is obvious that combined use with simultaneous administration such asin the form of a combination medicine or by means of directly successiveor temporally staggered administration of the various components canalso be made. As a matter of course the combination medicine as well asthe various components can be used in connection with pharmaceuticallyacceptable carriers or dilution agents.

According to yet another embodiment of the invention dosage can bespread out over at least two months and particularly preferable is overat least three months. However, this should only be understood as anexample and the concrete dosage duration can naturally be varied in eachindividual case. Optimum results are obtained by administering a workingdose of 12,000 units per day, five to seven times a week over a periodof six to eight weeks.

As an example, the cardiovascular disease or disorder may involve acoronary heart disease (CHD), all forms of arteriosclerosis, carotidstenosis, stenosis of the arteries leading to the brain cardiacinfarction, apoplexy, cardiac hypertrophy or fatty disease of the heart.

Additionally, the use of the enzyme for prevention of restenosis inpatients who have already been subjected to a coronary intervention isindicated. Coronary interventions include, for example, stentimplantation, balloon dilatation, rotablation and bypass operations(arterio-occlusive disease treatments).

It has moreover been found that according to the invention prevention ortreatment of acute cardiac infarction can be carried out.

Here below the invention is described in more detail without limitation.

Hyaluronidase is in itself well-known in the state-of-the-art andbelongs to the so-called β(1-4)-glycosidases. These enzymes are alsodesignated as hyaluronate glycan hydrolases, EC 3.2.1.35 through3.2.1.36. Hyaluronidase hydrolyses hyaluronic acid, a linearheteroglycan with alternating glucuronic acid and N-acetyl-glucosamineresidues (acidic glycosaminoglycan (mucopolysaccharide)) and hyaluronate(the ionic form of hyaluronic acid) but also does the same tochondroitin sulphate.

The hyaluronidase can be derived from any source whatsoever and, forinstance may be recovered from bovine protein (bovine type), leeches orbacteria (e.g. in the form of hyaluronate lyase). The hyaluronidase canalso be of vegetable origin. The hyaluronidase can be isolated, forinstance, from potatoes, tobaccos and peas. Genetic engineeringtechniques in the art can likewise be used to produce hyaluronidase.

Particularly preferred is any hyaluronidase which splits and thusdepolymerises hyaluronic acid, chondroitin-4-sulphate,chondroitin-6-sulphate and mucotin sulphate where the most preferredhyaluronidase is an enzyme available commercially such as, by way ofexample, the hyaluronidase marketed under the trade name of Hylase®“Dessau” by the firm of Pharma Dessau.

The hyaluronidase can of course not only be used to therapy an alreadyexisting disorder or disease of the cardiovascular system but also forpreventive treatment, that is prophylactically, of the indications citedabove in order to avoid or delay the occurrence of such disorders.

For treatment and/or prevention of the disorders indicated above, amixture of hyaluronidases of different origins can also be used.

Particularly well suited is any hyaluronidase for prevention in the caseof patients at risk of cardiac infarction and/or for treatment ofcoronary heart disease. Here the enzyme can also be administereddirectly intra-coronarily in addition to intravenous administration.

Coronary heart disease is also known under the terms of “stenoticcoronary sclerosis”, “degenerative coronary disease” and “ischaemicheart disease”. A pathophysiologic basis is reduction in blood supplycaused by sclerosis or occlusion of coronary vessels and thus the supplyof energy-providing substrate and oxygen to the heart muscle entailing adiscrepancy between supply and demand and clearly noticeable understrain (limited coronary reserves). As syndromes the following areknown; angina pectoris, cardiac infarction as well as left ventricularfailure. A CHD can also progress for a long time asymptomatically whilein the exercise ECG under certain conditions signs of a lack of bloodsupply can be shown.

The success of treatment can be checked, for example, by means of anECG, an exercise ECG, a long-term ECG, echocardiography, stress echo aswell as with myocardial scintigraphy and coronary angiography as well ascoronary angloscopy. It has generally been shown that by means ofadministration according to the present invention blood supply to thecoronary vessels was improved, to some extent even attaining completehealing.

The following examples will illustrate the invention without beinglimiting.

Numerous patients with cardiovascular diseases were treated, some ofwhom in particular suffered from coronary heart disease. The patientswere treated for at least four weeks, preferably for at least twomonths, and most preferably for at least three months, with ahyaluronidase-containing preparation, preferably with the medicamentHylase® “Dessau”. When administering 6,000 units per day of thishyaluronidase preparation, intravenously, by way of example, five times(up to seven times) per week, after several weeks a significantreduction in complaints was observed with the patients.

Some patients were also treated with dosages of Hylase® “Dessau” in therange of 12,000 units per day, five to seven times per week, and withdosages extending up to 15,000 units per day, five to seven times perweek. Besides the preferably used intravenous application, other kindsof application such as intramuscular or subcutaneous application, alsoled to success but where the intravenous application was the mosteffective method of administration.

When using other hyaluronidases than Hylase® “Dessau” other dosages maybe required which the person skilled in the art can easily determineaccording to the practical circumstances of the case.

According to another aspect of the invention, treatment ofcardiovascular diseases with hyaluronidase is done by way ofexemplification in combination with camitin. By using this combination,cardiac hypertrophy and fatty disease of the heart can be counteractedand thus the risk of an infarction can be significantly reduced.

Biochemically, the use according to the invention is directly connectedwith the degradation of fatty acids. Since the fatty acid synthesispreferably takes place in the cytoplasm while oxidative breakdown offatty acids takes place in the mitochondria, the fatty acids to bebroken down must first migrate from the cytoplasm to the mitochondria.Camitin (4-trimethylamino-3-hydroxybutyric acid) assists in suchmigration and can react both with acetyl CoA as well as with acyl CoAwith the formation of acetyl-carnitin or acyl-carnitin. The formation ofcamitin-activated acetic acid or fatty acid is catalysed by means ofacyl-CoA-camitin-transferase of which there are two distinct types (foracetyl transfer as well as for acyl transfer). By substituting theacetyl residue for a fatty acid residue a carnityl-fatty acid compoundis produced which can pass through the mitochondria membrane. Thisexplains the favourable effect of camitin on fatty acid oxidation. Afterentering the mitochondria, the fatty acid is transferred from theacylcarnitin to the CoA and the camitin is then again available forfatty acid transport.

Additional active ingredients neither possessing any coenzyme functionnor demonstrable as essential components for humans but beingindispensable cellular components and to some extent performingfunctions as yet not researched are collectively referred to asvitamin-like active agents. They to some extent influence thepermeability of membranes and capillaries.

These include, besides carnitin:

-   -   Meso-inositol (myo-inositol)    -   Essential fatty acids (vitamin F)    -   Flavanoids (vitamin P)

A further aspect of the invention relates to the use of hyaluronidase incombination with so-called CSE inhibitors (CSE=cholesterol synthesisingenzymes) such as pravasin and all other so-called “statins”. In thiscase it is unambiguously proven (e.g. in the WCS study) that pravasin inparticular and other statins as well have a cardio-protective effectAdditionally, they act against arteriosclerosis precisely ashyaluronidase does. The purpose of the combination is to achieveoptimisation of therapy by means of the two substances' synergies.

Additional substances which may favourably influence the effect ofhyaluronidase and which can be used in combinations according to theinvention:

-   1) Magnesium and other cations-   2) Nootropics (e.g. piracetam)-   3) Agents enhancing blood supply (e.g. pentoxifillin, ginkgo, etc.)-   4) Various vitamins (e.g. vitamin C)-   5) Substances increasing NO synthesis-   6) Glycoprotein antagonists such as eptifibatid-   7) Plasminogen activators such as altelase, retelase, etc. In case    of acute coronary thrombosis (cardiac infarction)

Use according to the invention is furthermore suitable for the followingindications:

-   1) Cerebral sclerosis-   2) Multiple sclerosis-   3) Oncology: improvement of resorption of cytostatics in diseased    tissue-   4) Ophthalmology (glaucoma)-   5) Diabetic mlcro-angiopathy-   6) Increase in fertility in men and women

1-6. (canceled)
 7. A method of preventing and/or treating cardiovasculardisease, which method comprises intravenously administering to a patientat least 6,000 units per day of hyaluronidase for at least four weeks,whereupon cardiovascular disease in the patient is treated and/orprevented.
 8. The method of claim 7, which method comprisesintravenously administering to the patient at least 12,000 units per dayof hyaluronidase.
 9. The method of claim 8, which method comprisesintravenously administering to the patient at least 15,000 units per dayof hyaluronidase.
 10. The method of claim 7, which method furthercomprises administering one or more agents selected from the groupconsisting of carnitin, a cholesterol synthesizing enzyme (CSE)inhibitor, a nootropic, a blood supply enhancer, a vitamin, and acation.
 11. The method of claim 8, which method further comprisesadministering one or more agents selected from the group consisting ofcarnitin, a CSE inhibitor, a nootropic, a blood supply enhancer, avitamin, and a cation.
 12. The method of claim 9, which method furthercomprises administering one or more agents selected from the groupconsisting of carnitin, a CSE inhibitor, a nootropic, a blood supplyenhancer, a vitamin, and a cation.
 13. The method of claim 7, whichmethod comprises administering hyaluronidase for at least two months.14. The method of claim 8, which method comprises administeringhyaluronidase for at least two months.
 15. The method of claim 9, whichmethod comprises administering hyaluronidase for at least two months.16. The method of claim 10, which method comprises administeringhyaluronidase and one or more other agents for at least two months. 17.The method of claim 11, which method comprises administeringhyaluronidase and one or more other agents for at least two months. 18.The method of claim 12, which method comprises administeringhyaluornidase and one or more other agents for at least two months. 19.The method of claim 7, wherein the cardiovascular disease involvescoronary heart disease, arteriosclerosis, carotid stenosis, stenosis ofarteries leading to the brain, cardiac infarction, apoplexy, cardiachypertrophy, or fatty disease of the heart.
 20. The method of claim 8,wherein the cardiovascular disease involves coronary heart disease,arteriosclerosis, carotid stenosis, stenosis of arteries leading to thebrain, cardiac infarction, apoplexy, cardiac hypertrophy, or fattydisease of the heart.
 21. The method of claim 9, wherein thecardiovascular disease involves coronary heart disease,arteriosclerosis, carotid stenosis, stenosis of arteries leading to thebrain, cardiac infarction, apoplexy, cardiac hypertrophy, or fattydisease of the heart.
 22. The method of claim 10, wherein thecardiovascular disease involves coronary heart disease,arteriosclerosis, carotid stenosis, stenosis of arteries leading to thebrain, cardiac infarction, apoplexy, cardiac hypertrophy, or fattydisease of the heart.
 23. The method of claim 11, wherein thecardiovascular disease involves coronary heart disease,arteriosclerosis, carotid stenosis, stenosis of arteries leading to thebrain, cardiac infarction, apoplexy, cardiac hypertrophy, or fattydisease of the heart.
 24. The method of claim 12, wherein thecardiovascular disease involves coronary heart disease,arteriosclerosis, carotid stenosis, stenosis of arteries leading to thebrain, cardiac infarction, apoplexy, cardiac hypertrophy, or fattydisease of the heart.
 25. The method of claim 13, wherein thecardiovascular disease involves coronary heart disease,arteriosclerosis, carotid stenosis, stenosis of arteries leading to thebrain, cardiac infarction, apoplexy, cardiac hypertrophy, or fattydisease of the heart.
 26. The method of claim 14, wherein thecardiovascular disease involves coronary heart disease,arteriosclerosis, carotid stenosis, stenosis of arteries leading to thebrain, cardiac infarction, apoplexy, cardiac hypertrophy, or fattydisease of the heart.
 27. The method of claim 15, wherein thecardiovascular disease involves coronary heart disease,arteriosclerosis, carotid stenosis, stenosis of arteries leading to thebrain, cardiac infarction, apoplexy, cardiac hypertrophy, or fattydisease of the heart.
 28. The method of claim 16, wherein thecardiovascular disease involves coronary heart disease,arteriosclerosis, carotid stenosis, stenosis of arteries leading to thebrain, cardiac infarction, apoplexy, cardiac hypertrophy, or fattydisease of the heart.
 29. The method of claim 17, wherein thecardiovascular disease involves coronary heart disease,arteriosclerosis, carotid stenosis, stenosis of arteries leading to thebrain, cardiac infarction, apoplexy, cardiac hypertrophy, or fattydisease of the heart.
 30. The method of claim 18, wherein thecardiovascular disease involves coronary heart disease,arteriosclerosis, carotid stenosis, stenosis of arteries leading to thebrain, cardiac infarction, apoplexy, cardiac hypertrophy, or fattydisease of the heart.
 31. A method of preventing and/or treatingcardiovascular disease, which method comprises administering to apatient at least 6,000 units per day of hyaluronidase for at least fourweeks whereupon cardiovascular disease in the patient is treated and/orprevented.